An Investigation of the Mineral in Ductile and Brittle Cortical Mouse Bone

Bone is a strong and tough material composed of apatite mineral, organic matter, and water. Changes in composition and organization of these building blocks affect bone's mechanical integrity. Skeletal disorders often affect bone's mineral phase, either by variations in the collagen or directly altering mineralization. The aim of the current study was to explore the differences in the mineral of brittle and ductile cortical bone at the mineral (nm) and tissue (µm) levels using two mouse phenotypes. Osteogenesis imperfecta model, oim‐/‐, mice have a defect in the collagen, which leads to brittle bone; PHOSPHO1 mutants, Phospho1‐/‐, have ductile bone resulting from altered mineralization. Oim‐/‐ and Phospho1‐/‐ were compared with their respective wild‐type controls. Femora were defatted and ground to powder to measure average mineral crystal size using X‐ray diffraction (XRD) and to monitor the bulk mineral to matrix ratio via thermogravimetric analysis (TGA). XRD scans were run after TGA for phase identification to assess the fractions of hydroxyapatite and β‐tricalcium phosphate. Tibiae were embedded to measure elastic properties with nanoindentation and the extent of mineralization with backscattered electron microscopy (BSE SEM). Results revealed that although both pathology models had extremely different whole‐bone mechanics, they both had smaller apatite crystals, lower bulk mineral to matrix ratio, and showed more thermal conversion to β‐tricalcium phosphate than their wild types, indicating deviations from stoichiometric hydroxyapatite in the original mineral. In contrast, the degree of mineralization of bone matrix was different for each strain: brittle oim‐/‐ were hypermineralized, whereas ductile Phospho1‐/‐ were hypomineralized. Despite differences in the mineralization, nanoscale alterations in the mineral were associated with reduced tissue elastic moduli in both pathologies. Results indicated that alterations from normal crystal size, composition, and structure are correlated with reduced mechanical integrity of bone. © 2014 The Authors. Journal of Bone and Mineral Research published by Wiley Periodicals, Inc. on behalf of the American Society for Bone and Mineral Research.

[1]  Jan L. Bruse,et al.  Reference point indentation is not indicative of whole mouse bone measures of stress intensity fracture toughness , 2014, Bone.

[2]  Joseph M. Wallace,et al.  Multi-scale analysis of bone chemistry, morphology and mechanics in the oim model of osteogenesis imperfecta , 2014, Connective tissue research.

[3]  R. Ritchie,et al.  How Tough Is Brittle Bone? Investigating Osteogenesis Imperfecta in Mouse Bone , 2014, Journal of bone and mineral research : the official journal of the American Society for Bone and Mineral Research.

[4]  Jennifer R. Melander,et al.  Gender-Dependence of Bone Structure and Properties in Adult Osteogenesis Imperfecta Murine Model , 2013, Annals of Biomedical Engineering.

[5]  Sandra J Shefelbine,et al.  Insight into differences in nanoindentation properties of bone. , 2013, Journal of the mechanical behavior of biomedical materials.

[6]  Noor Azuan Abu Osman,et al.  Morphological Change of Heat Treated Bovine Bone: A Comparative Study , 2012, Materials.

[7]  Kevin W Eliceiri,et al.  NIH Image to ImageJ: 25 years of image analysis , 2012, Nature Methods.

[8]  A. Boyde,et al.  Ultra-structural defects cause low bone matrix stiffness despite high mineralization in osteogenesis imperfecta mice☆ , 2012, Bone.

[9]  Alexander J. Makowski,et al.  The Contribution of the Extracellular Matrix to the Fracture Resistance of Bone , 2012, Current Osteoporosis Reports.

[10]  M. Tzaphlidou,et al.  Ca/P concentration ratio at different sites of normal and osteoporotic rabbit bones evaluated by Auger and energy dispersive X-ray spectroscopy , 2011, Journal of Biological Physics.

[11]  Vadim V. Silberschmidt,et al.  Micro-scale modelling of bovine cortical bone fracture: Analysis of crack propagation and microstructure using X-FEM , 2012 .

[12]  D. G. T. Strange,et al.  Size effects in indentation of hydrated biological tissues , 2012 .

[13]  Hrishikesh Bale,et al.  Age-related changes in the plasticity and toughness of human cortical bone at multiple length scales , 2011, Proceedings of the National Academy of Sciences.

[14]  M. Mello,et al.  Collagen type I amide I band infrared spectroscopy. , 2011, Micron.

[15]  I. Jasiuk,et al.  Multi-scale characterization of swine femoral cortical bone. , 2011, Journal of biomechanics.

[16]  D. Vashishth,et al.  The relative contributions of non-enzymatic glycation and cortical porosity on the fracture toughness of aging bone. , 2011, Journal of biomechanics.

[17]  M. McKee,et al.  Loss of Skeletal Mineralization by the Simultaneous Ablation of PHOSPHO1 and Alkaline Phosphatase Function: A Unified Model of the Mechanisms of Initiation of Skeletal Calcification , 2010, Journal of bone and mineral research : the official journal of the American Society for Bone and Mineral Research.

[18]  Simon R. Goodyear,et al.  PHOSPHO1 is essential for mechanically competent mineralization and the avoidance of spontaneous fractures. , 2010, Bone.

[19]  Y. Zuo,et al.  Comparative study on inorganic composition and crystallographic properties of cortical and cancellous bone. , 2010, Biomedical and environmental sciences : BES.

[20]  A. Boskey,et al.  Contribution of Mineral to Bone Structural Behavior and Tissue Mechanical Properties , 2010, Calcified Tissue International.

[21]  J. Millán,et al.  Inhibition of PHOSPHO1 activity results in impaired skeletal mineralization during limb development of the chick. , 2010, Bone.

[22]  Paul K. Hansma,et al.  Plasticity and toughness in bone , 2009 .

[23]  M. Tzaphlidou Bone Architecture: Collagen Structure and Calcium/Phosphorus Maps , 2008, Journal of biological physics.

[24]  A. Boyde,et al.  Composite bounds on the elastic modulus of bone. , 2008, Journal of biomechanics.

[25]  M. Buehler Nanomechanics of collagen fibrils under varying cross-link densities: atomistic and continuum studies. , 2008, Journal of the mechanical behavior of biomedical materials.

[26]  Timothy M. Wright,et al.  Abnormal Mineral-Matrix Interactions Are a Significant Contributor to Fragility in oim/oim Bone , 2007, Calcified Tissue International.

[27]  Markus J. Buehler,et al.  Molecular nanomechanics of nascent bone: fibrillar toughening by mineralization , 2007 .

[28]  M. Oyen Sensitivity of polymer nanoindentation creep measurements to experimental variables , 2007 .

[29]  J. Millán,et al.  Functional Involvement of PHOSPHO1 in Matrix Vesicle–Mediated Skeletal Mineralization , 2007, Journal of bone and mineral research : the official journal of the American Society for Bone and Mineral Research.

[30]  J. Kopp,et al.  Changes in apatite crystal size in bones of patients with osteogenesis imperfecta , 1991, Calcified Tissue International.

[31]  R. Legros,et al.  Age-related changes in mineral of rat and bovine cortical bone , 1987, Calcified Tissue International.

[32]  Michelle L. Oyen,et al.  Analytical techniques for indentation of viscoelastic materials , 2006 .

[33]  Joel W. Ager,et al.  Fracture and Ageing in Bone: Toughness and Structural Characterization , 2006 .

[34]  C. Farquharson,et al.  The presence of PHOSPHO1 in matrix vesicles and its developmental expression prior to skeletal mineralization. , 2006, Bone.

[35]  A. Boyde,et al.  Hydration effects on the micro-mechanical properties of bone , 2006 .

[36]  José M.F. Ferreira,et al.  Effect of Ca/P ratio of precursors on the formation of different calcium apatitic ceramics—An X-ray diffraction study , 2005 .

[37]  M. Grynpas,et al.  Age and disease-related changes in the mineral of bone , 2005, Calcified Tissue International.

[38]  Himadri S. Gupta,et al.  Structure and mechanical quality of the collagen–mineral nano-composite in bone , 2004 .

[39]  Michelle L. Oyen,et al.  Spherical indentation creep following ramp loading , 2005 .

[40]  F. Cui,et al.  Alterations in mineral properties of zebrafish skeletal bone induced by liliputdtc232 gene mutation , 2003 .

[41]  A. Boskey Bone mineral crystal size , 2003, Osteoporosis International.

[42]  A. Boskey,et al.  Infrared Analysis of the Mineral and Matrix in Bones of Osteonectin‐Null Mice and Their Wildtype Controls , 2003, Journal of bone and mineral research : the official journal of the American Society for Bone and Mineral Research.

[43]  K. Rogers,et al.  An X-ray diffraction study of the effects of heat treatment on bone mineral microstructure. , 2002, Biomaterials.

[44]  D. Bernache-Assollant,et al.  Calcium phosphate apatites with variable Ca/P atomic ratio II. Calcination and sintering. , 2002, Biomaterials.

[45]  P. Fratzl,et al.  Age- and genotype-dependence of bone material properties in the osteogenesis imperfecta murine model (oim). , 2001, Bone.

[46]  M. Epple,et al.  The structure of bone studied with synchrotron X-ray diffraction, X-ray absorption spectroscopy and thermal analysis , 2000 .

[47]  D. A. Bradley,et al.  Oim mice exhibit altered femur and incisor mineral composition and decreased bone mineral density. , 2000, Bone.

[48]  P. Sarathchandra,et al.  Abnormal Mineral Composition of Osteogenesis Imperfecta Bone as Determined by Electron Probe X-ray Microanalysis on Conventional and Cryosections , 1999, Calcified Tissue International.

[49]  A L Boskey,et al.  The Material Basis for Reduced Mechanical Properties in oim Mice Bones , 1999, Journal of bone and mineral research : the official journal of the American Society for Bone and Mineral Research.

[50]  P. Antich,et al.  Bone material elasticity in a murine model of osteogenesis imperfecta. , 1999, Connective tissue research.

[51]  D L Dorset,et al.  X-ray Diffraction: A Practical Approach , 1998, Microscopy and Microanalysis.

[52]  P Zioupos,et al.  Mechanical properties and the hierarchical structure of bone. , 1998, Medical engineering & physics.

[53]  P. Fratzl,et al.  Bone mineralization in an osteogenesis imperfecta mouse model studied by small-angle x-ray scattering. , 1996, The Journal of clinical investigation.

[54]  A L Boskey,et al.  Mineral changes in a mouse model of osteogenesis imperfecta detected by Fourier transform infrared microscopy. , 1996, Connective tissue research.

[55]  J. Clement,et al.  Age and temperature related changes to the ultrastructure and composition of human bone mineral , 1995, Journal of bone and mineral research : the official journal of the American Society for Bone and Mineral Research.

[56]  A. Boyde,et al.  Mineral density quantitation of the human cortical iliac crest by backscattered electron image analysis: variations with age, sex, and degree of osteoarthritis. , 1995, Bone.

[57]  M T Davisson,et al.  Defective pro alpha 2(I) collagen synthesis in a recessive mutation in mice: a model of human osteogenesis imperfecta. , 1993, Proceedings of the National Academy of Sciences of the United States of America.